Objective:To establish the rat cardiac arrest model in high-altitude hypobaric hypoxia environment, and to explore the effect of the treatment time in the hypobaric oxygen chamber on the reproduction of high-altitude rat cardiac arrest model.Methods:SPF grade healthy male Sprague-Dawley (SD) rats were used as observation subjects. The experiment was conducted in two different altitude areas. The rats from the Plateau Branch of Institute of Cardiopulmonary and Cerebral Resuscitation of Sun Yat-sen University (Xining, Qinghai) were weighed and numbered, and they were placed in a hypobaric oxygen chamber (simulated altitude of 3 000 meters, speed of ascent and descent of 15 m/min, temperature of 20 ℃, cabin pressure of 69.5 kPa, cabin oxygen pressure of 14.5 kPa). After 30 days of feeding, the rats were obtained according to random number table method, and the cardiac arrest model was established by asphyxia method as the 30-day hypobaric hypoxia group. After 60 days of feeding, rats were randomly selected again, and the cardiac arrest model was established as the 60-day hypobaric hypoxia group. Thirty rats were randomly selected from the Institute of Cardiopulmonary Cerebral Resuscitation at Sun Yat-sen University (Guangzhou, Guangdong) by the same method, and the cardiac arrest model was established as the plain control group. The differences in the body weight of rat modeling precursors and the induction time of asphyxia during the modeling process among different groups were compared.Results:Finally, cardiac arrest model was established in 16 rats in the 30-day hypobaric hypoxia group and in 22 rats in the 60-day hypobaric hypoxia group. There was no significant difference in the body weight of rats before modeling among the plain control group, 30-day hypobaric hypoxia group and 60-day hypobaric hypoxia group [g: 429.00 (389.25, 440.75), 440.00 (415.50, 486.25), 440.00 (400.00, 452.50), all P > 0.05]. The asphyxia induction time of rats in the 60-day hypobaric hypoxia group was significantly longer than that in the 30-day hypobaric hypoxia group (s: 294.59±75.39 vs. 234.31±93.86, P < 0.01), even about 1.4 times of the plain control group (s: 294.59±75.39 vs. 208.73±30.88, P < 0.01). There was no significant difference in the asphyxia induction time between the 30-day hypobaric hypoxia group and the plain control group ( P > 0.05). Conclusion:Rats treated in a hypobaric oxygen chamber for 60 days are more suitable for the preparation of high-altitude cardiac arrest model, and are also consistent with the oxygen reserve and hypoxia tolerance of high-altitude rats.

This study aims to investigate the function of lmo2363 gene in stress resistance of Liste-ria monocytogenes strain LM83-1.In this study,the lmo2363 gene deletion strain and complement-ation strain of Listeria monocytogenes were constructed using overlapping extended PCR and ho-mologous recombination techniques,and the growth ability,stress survival rate and biofilm forma-tion ability of wild,deletion strain and complementation strain were compared under different stress environments.lmo2363 gene deletion strain and complementation strain of Listeria monocy-togenes were successfully constructed in this experiment.The growth curves showed that the growth capacity of the deletion strain was weaker than the wild strain LM83-1 under 4 ℃,7％NaCl,10％NaCl,3.5％ethanol,4.0％ethanol and pH5 stress(P＜0.001).The results of stress survival test showed that the survival rate of the deletion strain was significantly lower than the wild strain after 1 h treatment with pH3 and 10 mmol/L H2 O2 stress(P＜0.010).The biofilm forming ability of the deletion strain was decreased compared with that of the wild strain(P＜0.050).This study confirmed that lmo2363 gene mediated the adaptation of LM to low temperature,high osmotic pressure,ethanol and acid stress environment and affected the formation of LM bio-film.This study laid a foundation for further exploring the function of lmo2363 gene in the stress resistance process of Listeria monocytogenes.

Objective: To investigate the expression of large tumor suppressor homolog 2 (LATS2) gene and its promotor methylation in oral squamous cell carcinoma (OSCC). Methods: Reverse transcription-PCR (RT-PCR) and pyrosequencing were used to detect the mRNA and promotor methylation of LATS2 gene in 72 OSCC specimens and normal oral mucosa tissues. Western blotting was used to detect the LATS2 protein in six OSCC specimens and normal oral mucosa tissues. Results: All cases had expression of LATS2 mRNA in normal oral mucosa tissues, but the expression was down-regulated significantly, only 47% (34/72) in 72 cases of OSCC showed LATS2 mRNA expression. The expression was correlated with the degree of tumor differentiation and lymph node metastasis (P<0.05). The results of pyrosequencing show that 68% of promotor methylation (49/72) in 72 cases of OSCC. Furthermore, there was significant correlation between the mRNA and promotor methylation of LATS2 gene (χ²=16.980, P<0.01). All the six specimens had the low LATS2 protein expression. Conclusions The promotor methylation of LATS2 gene may play an important role in the occurrence of OSCC.

RT-PCR and Western blot were used to detect the LATS2 mRNA and protein level in 57 cases of oral squamous cell carcinoma (OSCC)specimens and 12 normal oral mucosa tissues.LATS2 mRNA and protein were expressed in all the normal oral mucosa samples (100%),but only 47.4% (27 /57)and 42.1% (24 /57)in OSCC samples respectively with positive correlation of both(P <0.01),and were respectively correlated with the tumor differentiation degree and lymph node metastasis(P <0.05).

OBJECTIVETo apply single nucleotide polymorphism (SNP) microarray for delineation of small supernumerary marker chromosome (sSMC) in two newborns.

METHODChromosome karyotyping was performed on newborns who were born in Jan. 2013 and Jan. 2014 in Haidian Maternal and Child Health Hospital because of the abnormalities found in pregnancy checkups. SNP microarray analysis was carried out on 2 newborns with de novo sSMCs (one was mos 47,XY, + mar[45]/46,XY[5] and the other was mos 47, XY, + mar [30]/46, XY [20]), which could not be determined by conventional banding techniques. Genomic DNA was extracted from cord blood samples, amplified, tagged and hybridized following the manufacturer' s protocol. Data were collected and analyzed.

RESULTThere was a 78. 6 Mb duplication in chromosome 8 for Newborn A, which was associated with 8p22 duplication syndrome; and a 32. 7 Mb duplication in chromosome 13 for Newborn B, which was not yet reported definitely as pathogenic. The newborn A was identified with agenesis of the corpus callosum, obvious right eyelid drooping, the onset of low muscle tone and mental developmental lag behind their peers, while the newborn B had normal findings on physical and mental evaluation.

CONCLUSIONSNP-array can identify sSMCs of newborns at the DNA level, and can be used as an important supplement to the conventional karyotype analysis, but the pathogenicity of positive outputs need further verification.

Chromosome Duplication ; Chromosomes, Human, Pair 8 ; Genetic Markers ; Humans ; In Situ Hybridization, Fluorescence ; Infant, Newborn ; Karyotyping ; Oligonucleotide Array Sequence Analysis ; Polymorphism, Single Nucleotide

Objective To explore the feasibility of inter-laboratory comparison and trueness evaluation among clinical laboratories, and assess the quality of participants′measurement, by measuring the activity of alanine aminotransferase ( ALT ) in patient serum samples.Methods Method comparison study was used.Five patients serum samples, whose target values were assigned by two international candidate reference laboratory with reference method of ALT without pyridoxal phosphate, were measured by 23 routine laboratories.The bias between measurement result of each participant and the mean of reference laboratories was calculated, and then compared to allowable bias 6%.Calculate the mean value and the relative bias.Results Compared with the mean of reference laboratories, the maximum absolute value of bias among the 23 routine laboratories was 31.27%.The rate range which bias was less than the allowable bias was 26.09%-73.91 %.The bias acceptability of 8 participants were more than or equal to 80%;15 participants were less than or equal to 60%; and 3 participants were 0%.Conclusions Using patient serum samples and values assigned by reference method is an effective way to carry out inter-laboratory comparison and trueness evaluation.It can reflect the quality of measurement more truly.

OBJECTIVETo investigate the clinicopathological characteristics and the diagnosis of multilocular cystic renal cell carcinoma (MCRCC).

METHODSThe clinicopathological data of 19 MCRCC cases were collected and immunohistochemical staining assays were carried out. Forty-six cases of other cystic kidney lesions within the same period were collected as controls, including extensively cystic clear cell RCC (12 cases), clear cell tubulopapillary renal cell carcinoma (6 cases), tubulocystic carcinoma (2 cases), simple cortical cysts (22 cases), multilocular cystic nephroma (1 cases) and multicystic kidney (3 cases).

RESULTSThe patients included 14 males and 5 females. The ages ranged from 31 to 66 years (median age = 50 years). Most of the MCRCC cases were detected incidentally in physical examination, occasionally accompanied with hematuria, back pain or other symptoms. The follow-up period of 17 patients ranged from 6 to 170 months. All patients were alive without evidence of tumor recurrence or metastasis. Pathological findings showed that macroscopically, tumor size ranges from 1.5 to 7.0 cm in the maximum diameter, generally a entirely of various sized. The cysts contain serous, hemorrhagic or turbid fluid. Solid areas or substantially discernible mural nodules were absent; histologicallly, single layer of cuboidal and flattened epithelial tumor cells were lined in the cysts, described as clear cytoplasm, small nuclear, no nucleoli and low Fuhrman nuclear grade (I or II). Multilayer tumor cells could be observed in a few cysts, with granular cytoplasm and small intracystic papillae formed. The clear tumor cell clusters, similar as cystic lined tumor cells, were seen within pathological fibrous in almost all cases, and significant myofibroblastic proliferation was found in 14 cases. Immunohistochemically, the cysts lined epithelial cells and the clear tumor cell clusters were positive for epithelium markers, including CKpan(19/19), EMA(16/19) and CK7 (15/19); higher percentage of CAIX (17/19) and PAX8(15/19) than control groups, but lower percentage of CD10 (7/19), RCC (6/19) and AMACR(2/19); and all were negative for 34βE12, CD117 and CD68.

CONCLUSIONSMultilocular cysts, clear cells clusters of low Fuhrman grade within fibrous septa and capillary vessel proliferation under epithelium are important features of MCRCC. The united using of CAIX, CK7, CD10 and RCC is helpful for differentiating variable cystic renal tumor. MCRCC usually has an excellent prognosis, nephron sparing surgery is first recommended as a therapeutic strategy.

Adenocarcinoma, Clear Cell ; metabolism ; pathology ; Biomarkers ; Carcinoma, Renal Cell ; metabolism ; pathology ; Cysts ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Humans ; Kidney Diseases, Cystic ; metabolism ; pathology ; Kidney Neoplasms ; metabolism ; pathology ; Male ; Neoplasm Recurrence, Local ; Prognosis ; Racemases and Epimerases ; metabolism

Objective:To study the serological reactions in cynomolgus monkeys infected with hepatitis B virus ( HBV). Methods:To select 1 to 3 days old or adult healthy cynomolgus monkeys by artificial breeding to observe the virology screening in laboratory a month to confirmed healthy animals ,randomly divided into control group and infection group .Infection group vaccination serum HBV carriers 0.5 ml (HBV-DNA≥108 copies) single cages,observe each group behavioral changes daily after inoculation 1 to 12 weeks, each week to confirmed the degree of liver inflammation through the HBV-M, HBV-DNA, liver function and on the B-guided, liver tissue inflammation by routine HE staining .Results: Adult monkeys did not induce positive reaction after vaccination , there were three young monkeys appear HBsAg , HBcAb and 2 appear HBV-DNA reaction, ALT poison attack occurred in HBsAg-positive began to increase after one week , one month after the peak , which was 180 U/L, after gradually decreased , continuing a month later near normal .AST higher than a week after the normal reference values were flat curve , representing the peak ALT after a month later, HBsAg positive cynomolgus monkeys HE staining showed mild hepatitis partial liver tissue lesions .Conclusion:HBV-M, HBV-DNA, ALT, AST and liver histopathology after HBV infection have changed , this result showing that it's produce inflammation and induction the response of immune .

Objective To investigate the clinicopathological characteristics and the diagnosis of multilocular cystic renal cell carcinoma ( MCRCC).Methods The clinicopathological data of 19 MCRCC cases were collected and immunohistochemical staining assays were carried out .Forty-six cases of other cystic kidney lesions within the same period were collected as controls , including extensively cystic clear cell RCC (12 cases), clear cell tubulopapillary renal cell carcinoma (6 cases), tubulocystic carcinoma (2 cases), simple cortical cysts (22 cases), multilocular cystic nephroma (1 cases) and multicystic kidney (3 cases). Results The patients included 14 males and 5 females.The ages ranged from 31 to 66 years ( median age=50 years ).Most of the MCRCC cases were detected incidentally in physical examination , occasionally accompanied with hematuria , back pain or other symptoms.The follow-up period of 17 patients ranged from 6 to 170 months.All patients were alive without evidence of tumor recurrence or metastasis.Pathological findings showed that macroscopically , tumor size ranges from 1.5 to 7.0 cm in the maximum diameter , generally a entirely of various sized.The cysts contain serous , hemorrhagic or turbid fluid.Solid areas or substantially discernible mural nodules were absent; histologicallly , single layer of cuboidal and flattened epithelial tumor cells were lined in the cysts , described as clear cytoplasm , small nuclear , no nucleoli and low Fuhrman nuclear grade ( I or II).Multilayer tumor cells could be observed in a few cysts , with granular cytoplasm and small intracystic papillae formed.The clear tumor cell clusters , similar as cystic lined tumor cells, were seen within pathological fibrous in almost all cases , and significant myofibroblastic proliferation was found in 14 cases.Immunohistochemically , the cysts lined epithelial cells and the clear tumor cell clusters were positive for epithelium markers , including CKpan(19/19), EMA(16/19) and CK7 (15/19);higher percentage of CAⅨ(17/19)and PAX8(15/19) than control groups, but lower percentage of CD10 (7/19), RCC (6/19) and AMACR (2/19); and all were negative for 34βE12, CD117 and CD68.Conclusions Multilocular cysts , clear cells clusters of low Fuhrman grade within fibrous septa and capillary vessel proliferation under epithelium are important features of MCRCC.The united using of CAⅨ, CK7, CD10 and RCC is helpful for differentiating variable cystic renal tumor.MCRCC usually has an excellent prognosis, nephron sparing surgery is first recommended as a therapeutic strategy.

ObjectiveTo explore the feasibility and outcomes of natural orifice transanal laparoscopic Soave procedure for Hirschsprung's disease and allied disorders (HAD).MethodsFrom March 2010 to December 2011,31 cases (at the age from 3 mos to 6 yrs) with Hirschsprung's disease or allied disorders (5 cases)underwent laparoscopic-assisted Soave pull-through procedure at two tertiary hospitals.We modified this technique by mobilizing the left hemicolon or whole colon via rectal muscular sleeve approach under transanal or transumbilical laparoscopic vision,then endorectal pull-through to completearectosigmoidectomyorsubtotalcolectomy. ResultsAllprocedureswerecompleted successfully.A rectosigmoidectomy was performed in 16 cases with classic HD and subtotal colectomy in 15 cases with extended HD and HAD.The average operative time was ( 117 ± 13) min.The length of the resected segment was 35 -80 cm,and the estimated blood loss was 5 -20 ml. One infant developed postoperative intestinal obstruction that required open exploration.Follow-up of one to 20 mos found no stoma stenosis or constipation recurrence. Enterocolitis developed in 1patient.ConclusionsTransanal or transumbilical laparoscopic-assisted Soave pull-through surgery is safe,effective and with a benefit of much less invasion and almost invisible scars.